摘要
目的探讨瘤体注射平内阳霉素对增殖性血管瘤血管内皮细胞生长因子(VEGF)、碱性成纤维细胞长因子(bFGF)和增殖细胞核抗原(PCNA)表达水平的影响及可能机制。方法增殖血管瘤140例随机分成7组,每组20例,A组作为对照组;B组平阳霉素(1mg/ cm2)术前1周瘤体注射;C组平阳霉素加地塞米松术前1周瘤体注射; D组平阳霉素术前2周瘤体注射;E组平阳霉素加地塞米松术前2周瘤体注射; F组平阳霉素术前3周瘤体注射;G组平阳霉素加地塞米松术前3周瘤体注射。对收集到的血管瘤标本采用组织化学方法和免疫组织化学方法(SP法),进行光镜下组织结构观察及VEGF、PCNA和bFGF的含量检测。结果A、F、G组VEGF的表达水平为78.00±3.57,75.90±2.63,70.54±3.11,B、C、D、E组为56.35±2.60,41.50±2.12,40.10±2.27,27.70±1.40差异有显著性(P<0.05)。A、B、F、G组bFGF的表达水平81.40±2.00,73.55±1.67, 71.45±3.88,69.70±3.76, C、D、E组53.65±2.29,45.35±1.82,31.90±2.55,差异有显著性(P<0.05)。A、F、G组PCNA的表达水平为79.55±3.15, 76.30±3.71,69.20±2.97, B、C、D、E组57.30±2.34,44.95±2.47,44.25±1.60,34.00±2.25,差异有显著性(P<0.05)。结论平阳霉素瘤体注射可明显抑制血管瘤组织中VEGF和bFGF的表达,从而抑制其增殖。平阳霉素与糖皮质激素(地塞米松)联合应用局部注射能起到协同作用,术前2周局部注射平阳霉素或联合激素可缩小血管瘤的体积,减少术中出血,减小术后创面。
Objective To explore the influence of intratumoral injection of pinyangmycin on the expression levels of VEGF,PCNA and bFGF in proliferating hemangioma. methods One hundred and forty patients with proliferating hemangioma were randomly divided into seven groups, twenty patients every group .Group A as control group. Group B were injected with Pinyangmycin(1mg|ml) before 1 week .Group C were injected with pinyangmycin and dexamethasonum before one week. Group D were injected with Pinyangmycin before two weeks. Group E were injected with pinyangmycin and dexamethasonum before two weeks. Group F were injected with Pinyangmycin before three weeks. Group G were injected with pinyangmycin and dexamethasonum before three weeks. Observe the tissue configuration of above examples by histochemistry method and detect the expression level of the VEGF,bFGF and PCNA by immunohistochemistry method. Results The expressions of VEGF in the A,F,G groups were 78.00±3.57,75.90±2.63,70.54±3.11 and in the B,C,D,E groups were 56.35±2.60,41.50±2.12,40.10±2.27,27.70±1.40.The variance was significance(P<0.05). The expressions of bFGF in the A,B,F,G groups were 81.40±2.00,73.55±1.67, 71.45±3.88,69.70±3.76 and in the C,D,E groups were 53.65±2.29,45.35±1.82,31.90±2.55, The variance was significance(P<0.05). The expressions of PCNA in the A,F,G groups were 79.55±3.15, 76.30±3.71,69.20±2.97 and in the B,C,D,E groups were 57.30±2.34,44.95±2.47,44.25±1.60,34.00±2.25 The variance was significance(P<0.05). Conclusion Intratumoral injection of pinyangmycin in proliferating hemangioma may obviously restrain the expressions of VEGF and bFGF to inhibit it proliferation. Dexamethasonum and Pinyangmycin enhance role of each other . intratumoral injection of Pinyangmycin and Dexamethasonum before operating will shrink area of hemangioma.decrease demage of operation to body.
引文
1.王延安,蒋欣泉,郑家伟,张志愿,等,血管瘤发病机制的研究进展[J],中国口腔颌面外科杂志,2005,3(3):76—79.
2.Tan S T,Velickovic M,Ruger B M,et al.Cellular and extracellular markers ofhemangioma[J].plast.2000,106(3):529—538.
3.林晓曦,王炜,曹谊林.血管瘤和血管畸形治疗中的美容原则探讨[ J ].中华医学美学美容杂志, 2003, 9 (3) : 190 - 192.
4.曹东升,盛辉,张林,等.婴幼儿血管瘤的阶段性治疗[J].实用儿科临床杂志,2007,22(11)815—16.
5.盛辉,曹东升,张林,等.婴幼儿血管瘤的综合治疗[J].安徽医科大学学报,2006,41(6):709—10.
6.O’Keefe M , Lanigan B , Byrne S A. Capillary haemangioma of t heeyelids and orbit : a clinicalreview of t he safety and efficacy of intralesional steroid. Acta Opht halmol Scand , 2003 ,81 :294-98.
7.张斌,刘毅.体表血管瘤的药物治疗[J].中国综合临床,2003,19(5):389-390.
8.刘学键,秦中平,李克雷.平阳霉素治疗口腔颌面部血管瘤的远期疗效及适应证探讨[J].上海口腔医学,2001,10(4):295-298.
9.朱晓海,江华,袁湘斌,等.平阳霉素瘤内注射治疗体表血管瘤(附25例报告)[J].第二军医大学学报,2005,26(1):51-52.
10.陈瑞卿.平阳霉素注射治疗血管瘤150例疗效观察[J].现代中西医结合杂志,2004,13(14):1853.
11.Takahashi K,Mulliken J B,Kozakewich H P,et al The cellular markers that distinguish the phase of hemangiomas during infancy and childhood[J]. J Clin Invest,1994,93(6):2357—64.
12.Chang J,Most D,Bresnick S,et al.Proliferative hemangiomas:anslysis of cytoline gene expression and angiogenesis[J].Plast Reconst Surg.1999,103(1):1—9.
13.Hell P A,Lerison D A,Woods A L,et al.Proliferating cell nuclear antigen(PCNA) imnunolocailization in paratlin sections an index of cell proliferation with evidence of deregula tell expression in some neoplasms[J].Pathol,1990,162:285—89.
14.Mulliken JB , Glowacki J . Hemangiomas and vascular malformations in infant s and children : a classification based on endot helialcharacteristic. Plast Reconst r Surg , 1982 ,69 :4122422.
15.郑勤田,伍连康,容文星,等.平阳霉素局部注射治疗小儿血管瘤[J].中华外科杂志,1991,29(5):290—91.
16.胡文铎,崔乃杰,高仲阳.国家基本药物及新特药临床指南[M].津:天津科技翻译出版公司,1996:407-408.
17.Teighner R J,Povirk L F.Bleomycin-induced DNA lesions at mutalional hot spots:Implications for the mechanism of double-strand cleavage[J].Proc Nall Aced Sci USA,1990,87:8350.
18.彭心昭,沈忠英,黎妙娟,等.平阳霉素诱导人类食管癌细胞EC8712凋亡的研究[J].癌症,1999,l8(6):657-660.
19.Tai KW, Chou MY, Hu CC, et a1.Induction of apoptosis in KB cells by Pingyangmycin[J].Oral Oncol,2000,36(2):242-247.
20.陈传俊,张志愿.平阳霉素抑制舌鳞癌细胞系的量效和时效关系及可能的抗癌机制[J].中国口腔颌面外科杂志,2004,12,2(4):261-265.
21.李平,周水淼,李兆基,等。平阳霉素对大鼠脾脏组织的作用及其机制[J].第二军医大学学报,2006,2,27(2):178—181.
22.曾爱荣。自由基在平阳霉素硬化治疗血管瘤中的作用[J].临床医药实践杂志2003 ,8 ,12(8):579—580.
23.郑翔,赵怡芳.平阳霉素治疗后血管瘤自由基的检测[J ] .口腔面外科杂志,2001 ,2 :109.
24.单国平.平阳霉素治疗颌面部海绵状血管瘤的临床分析—附102例报告[J ] .口腔颌面外科杂志,2001 ,1 :88.
25.Weiss SJ , Young J ,Lobuglio AF et al . Role of hydrogenperoxide innentrophil mediated destruction of cultured endothelial cells[J ] . J Clin Invest ,1981 ,68 :714.
26. Fanton J ,Ward PA. Role of oxygen derived free radicalsand metabolites in leukocyte dependent inflammatory reactions[J ] . Am J Pathos ,1982 ,107 :397.
27.陈光,成继民,孙辉臣,等.颌面部血管瘤畸形动脉栓塞术后的病理学变化[J ] .实用口腔医学杂志,1999 , 15 :433.
28.吴洪斌.平阳霉素的不良反应[J].药物不良反应杂志,2000,4:245-246.
29.张永宽,葛前进,王兴强.平阳霉素治疗颌面部血管瘤常见的局部并发症分析[J].中国美容医学,2004,l3(5):554.
30.刘群英.平阳霉素+地塞米松局部注射治疗海绵状血管瘤35例临床分析[J].中国全科医学,2004,7(11):835.
31.Tokunaga T , Oshika Y, Abe Y, et al . Vascular endothelial growth factor (VEGF) mRNA isoform expression pattern is correlated with livermetastasis and poor prognosis in colon cancer. Br J Cancer , 1998 ,77 :998--1002.
32.Campbell C L , Savarese D M, Quesenberry PJ , et al . Expression ofmultiple angiogenic cytokines in cultured normal human prostate epithelial cells :predominance of vascular endothelial growth factor. Int J Cancer , 1999 ,80 :868--874.
33.Taichman N , Young S , Cruchley A , et al . Human neutrophils secretevascular endothelial growth factor[J]. Leukocyte Biol ,1997 ,62 :397--400.
34. Alvarez Arroyo MV , Caramelo C , Angeles Castilla M, et al . Role ofvascular endothelial growth factor in the response to vessel injury. Kidney Int Suppl ,1998 ,68 : 729.
35.Metais C , Li JY, Li J , et al . Effects of coronary artery disease on expression and microvascular response to VEGF. Am J Physiol , 1998275 :H1411--H1418.
36 . Ferrara N. Vascular endothelial growth factor. Eur J Cancer , 1996 ,32A:2413--2422.
37.Peters MC , Isenberg BC , Rowley JA , et al . Release from alginate enhances the biological activity of vascular endothelial growth factor. J Biomater Sci Polym Ed , 1998 ,9 :1267--1278.
38.Nor JE , Christensen J , Mooney DJ , et al . Vascular endothelial growthfactor (VEGF)-mediated angiogenesis is associated with enhanced endothelial cell survival and induction of Bcl22 expression. Am J Pathol ,1999 ,154 :375--384.
39.Bousvaros A , Leichtner A , Zurakowski D , et al . Elevated serum vascular endothelial growth factor in children and young adults with Crohn’s disease. Dig Dis Sci , 1999 ,44 :424--430.
40.Bates D , Curry F. Vascular endothelial growth factor increase microvascular permeability via a calcium dependent pathway. Am J Physiol ,1977 ,273 :H687--H694.
41 . Sommer A,Brewer MT,Thompson RC,et al.biochem Biophys Res Commun,1987,144(3)543—550.
42 . Okada-Ban M,Thiery JP,Jouanneau J.Int J Biochem Cell Biol,2000,32(3);263—267.
43.Prelich G,,Tan CK,Kostura M,et al.Functional identity of proliferating cell nuclear antigen and a DNA Polynerasedelta zuxiliary protein[J].Nature,1987,326(4):517—610.
44.Hell PA,Lerison DA,Woods AL,et al.Proliferating cell nuclear antigen(PCNA) imnunolocailization in paratlin sections an index of cell proliferation with evidence of deregula tell expression in some neoplasms[J].Pathol,1990,162:285—289.
45.俞松,刘文英,等。糖皮质激素治疗血管瘤的试验研究[J].中华医学杂志,2005,85(16):1121—1124.
46.野战涛,牛爱国,张玉斌,等。平阳霉素乳剂对增殖期毛细血管瘤组织中VEGF和bFGF的影响[J].河北医药,2007,29(1)44—46.
47.Seghezzi G,Patel S,Ren CJ,et al.Fibroblast growth factor-2(FGF-2) Induces vascular endothelial growth factor(VEGF) expression in the endothelial cells of forming capillaries:An autocrine mechanism contributing to angiogenesis[J].Journal of Cell Biology,1998,141(7);1659.
48.刘潜,李静,周德凯,等.糖皮质激素瘤内注射治疗婴幼儿血管瘤机理的研究[J].第四军医大学学报,2007,28(13):1199—1201.
49.刘文修。大剂量强的松冲击疗法治疗婴幼儿血管瘤25例报告[J].华西口腔医学杂志,1993,11(1):59—60.
1.王延安,蒋欣泉,郑家伟,张志愿,等,血管瘤发病机制的研究进展[J],中国口腔颌面外科杂志,2005,3(3):76—79.
2.Tan ST,Velickovic M,Ruger BM,et al.Cellular and extracellular markers of hemangioma[J].plast,2000,106(3)529—538.
3. Iwata J , Sonobe H , Furihata M , et al . High f requency of apoptosis in infantile capillary hemangioma. J Pat hol , 1996 , 179 ( 4) :403～408.
4. Razon MJ , Kraling BM , Mulliken JB , et al . Increased apoptosiscoincides wit h onset of involution in infantile hemangioma. Micro-circulation , 1998 , 5 (223) : 189～195.
5.程立新,梁杰,汤少明,等.细胞凋亡抑制基因bcl-2在血管瘤发病中作用的研究.中华整形烧伤外科杂志,1999 ,5 (1) :35～36.
6.Gatalica Z,Lele SM, Rampy BA, et al. The exp ression of Fhit p rotein isrelated inversely to disease p rogression in patientswith breast carcinoma[ J ]. Cancer, 2000, 88 (6) : 1378 - 1383.
7.陈超,杨体泉。婴幼儿血管瘤内皮细胞凋亡的研究进展。实用儿科临床杂杂志,2007,6(22):869—870.
8.李静岩. Caspase-3与肿瘤关系的研究[ J ].医学综述, 2005, 11 ( 5) :430 - 432.
9. Hishikawa K,Nakaki T, Fujii T. Connective tissue growth factor induceapop tosisvia to caspase - 3 in cultural human cortic smooth muscle cells[ J ]. Eur J Pharm acol, 2000, 392 (1 - 2) : 19 - 22.
10.陈志雄,张瑞莲,陕声国,等. Caspase23在皮肤血管瘤不同时期的表达及意义[ J ].中华试验外科杂志, 2004, 21 (9) : 1109 - 1110.
11.李南,陈辛生,陈大良. Caspase23在血管瘤组织中的表达及其意义[ J ].中国普外基础与临床杂志, 2004, 11 (4) : 311 - 313.
12. Yang LQ, Fang DC,Wang RQ, et al. Effect of NF - kappaB, survivin,Bcl - 2 and Caspase 3 on apop tosis of gastric caner cells induced bytumor necrosis factor related apop tosis inducing ligand[ J ]. W orld J Gas2troenterol, 2004, 10 (1) : 22 - 25.
13. Suzuki A, Ito T, Kawano H, et al. Survivin initiate procaspase 3 /p21complex formation as a result of interaction with CD IS4 to resist fasme2diated cell death[ J ]. Oncogene, 2000, 19 (10) : 1346 - 1353.
14. Vegran F, Boidot R, Ondin C, et al. Distinct exp ression of Survivinsp lice variants in breast carcinomas [ J ]. Int J Oncol, 2005, 27 ( 4 ) :1151 - 1157.
15.Li YH,Wang C,Meng K, et al. Influence of survivin and caspase -3 oncell apop tosis and prognosis in gastric carcinoma[ J ]. World J Gastroen2terol, 2004, 10 (13) : 1984– 1988.
16.张文强,陈言汤,瞿晓梅. Survivin、TRA L在血管瘤中的表达[ J ].河南肿瘤学杂志, 2005, 18 (1) : 11 - 13.
17.钱江,乔成钢,柳宏,等。婴幼儿血管瘤组织中survivin和caspase -3的表达及其与细胞凋亡的关系[J].山东医药,2008,12(48):7—9.
18. Oda E , Ohki R , Murasawa H , et al . Noxa , a BH32only mem2ber of t he Bcl22 family and candidate mediator of p53 induced apop2tosis. Science ,2000 ,288 :1053～1058.
19.Gadd SH , Hobbs G, Miller MR , et al . Acetaminophen inducedproliferation of est rogen responsive breast cancer cells is associatedwit h increasesin cmyc RNA expression and NF2kappa b activity.Toxicol Sci , 2002 , 66 :233～2391.
20. Pinton P, Ferrari D, Rap izzi E, et al. The Ca2 + concentration of the endop lasmic reticulum is a key determinant of ceramide - induced apoptosis: Significance for the molecular mechanism of Bcl - 2 action [ J ].EMBO J , 2001, 20 (11) : 2690– 2701.
21. Kuwana T,Bouchier - Hayes L, Chipuk JE, et al. BH3 domains of BH3only proteoms differerntially regulate Bax - mediated mitochondricalmembranne pemeabilizahilization both directly and indirectly[ J ]. M olCell, 2005, 17 (4) : 525– 535.
22.李辉,张端莲,陕声国。bcl-2和bad蛋白表达与皮肤血管瘤的相关性研究[J]。数理医药学杂志,2007,2(20):160—163.
23.黄莺,王康敏,李恭才,等.婴幼儿血管瘤组织中Bcl-2和Bax与细胞凋亡的研究[ J ].中华小儿外科杂志, 2005, 26 (3) : 116 - 118.
24. Mancini AJ, Smoller BR. Proliferation and apop tosis within juvenilecap illary hemangiomas[ J ]. Am J Derm atopathol, 1996, 18 ( 5) : 505 -514.
25.王彪,庄福连,张鹏飞,等。细胞凋亡及bcl-2、Bax在血管瘤和血管畸形中的表达[J]。中华整形外科杂志,2003,9(5):347—349.
26. Yin XM,Oltval ZN, Korsmeyer SJ, et al. BH1 and BH2 domains of bcl-2 are required for inhibition of apop tosis and heterodimerizationwith Bax. Nature, 1994, 369 (6478) : 321--323.
27.BarinageM. Cell suicide: By ice, not fire. Science, 1994, 263: 754—756 .28.陈光,贺占国,成继民,等.颌面部毛细血管瘤中凋亡相关癌基因蛋白的表达.华北国防医药,2002 ,14 :79-80.
29.李辉,张端莲,陕声国。促凋亡基因Bad在皮肤血管瘤组织中的表达[J]。数理医药学杂志,2007,20(1):10—12.
30. Ottilie S , Diaz JL , Horne W, et al . Dimerization properties ofhuman BAD. Identification of a BH-3 domain and analysis of it sbinding to mutant bcl-2 and bcl-xL proteins. J Biol Chem ,1997 ,272 (49)∶30866.
31. Kelekar A , Chang BS , Harlan J E , et al . Bad is a BH23 domaincontaining protein t hat forms an inactivating dimer wit h bcl - xL.Mol Cell Biol , 1997 ,17 (12)∶7040.
32. Schimmer AD , Hedley , Pham NA , et al . BAD induces apoptosisin cells overexpressing Bcl-2 or Bcl-xL wit hout loss of mitochondral membrane potential . Leuk Lymphoma , 2001 , 42 (3)∶429.
33.Hattori T , Ookawa N , Fujita R , et al . Heterodimerization ofbcl-2 and bcl-x (L) wit h bax and bad in colorectal cancer . ActaOncol ,2000 ,39 (4)∶495.
34.韩志辉,薛振恂. Fas抗原在血管瘤及血管畸形中的表达及意义.实用口腔医学杂志,2002 ,18 :316-318.
35.邢新,袁斯明,欧阳天祥,等。Fas/FasL在各阶段婴幼儿血管瘤中的表达及意义[J]。中华整形外科杂志,2006,3(2):116—120.
36.陈光,贺占国,王琪,等.毛细血管组织中免疫细胞的分布变化及其意义.口腔医学,2000 ,20 :14-16.
37. Jia L , Patwari Y, Kelsey SM,et al . TRAIL induced apoptosis in typeⅠleykemic cells is not ten hanced by over expression of bax[J ] . BilchemBiophys Res Commun ,2001 ,283 :1037-1045.
38. Robert MP , Scot AM, Siegfrie DR , et al . Induction of apoptosis by Apo-Ligand , a new memberof the tumor necrosis factor cytosine family [J ] .Bi2ol Chem,1993 ,271 :12678.
39.王琪影,牛扶幼,翟晓梅。小儿血管瘤组织蛋白与细胞增殖和凋亡检测[J]。郑州大学学报,2007,5(3):548—550.
40. McMillan JR, McGrath JA, Pulkkinen L, et al. Br J Dermatol,1997, 136(6):817- 822.
41.盛迎伦,郑庆璇,赵晖,等。血管瘤和血管畸形病变组织中肥大细胞特点观察。中华病理学杂志,1994,23:311—313.
42. Pasyk KA, Argenta LC, Austad ED. Clin Plast Surg,1987, 14(3):435- 445.
43. Takahashi K, Mulliken JB, Kozakewich HP, et a1. J ClinInvest, 1994, 93(6):2357-2364.
44.Tan ST, Hasan Q, Velickovic M, et a1. Mod Pathol,2000, 13(1):92- 99.
45. Sgonc R , Fuerhapter C , Boeck G, et al. Induction of apoptosisin human dermal microvascular endothelial cells and infantile hemangiomas by interferon - alpha[J ] . Int Arch Allergy Immunol ,1998 , 117 (3) : 209.
46.Chang J , Most D , Bresnick S , et al. Proliferative hemangiomas :analysis of cytokine gene expression and angiogenesis [ J ] . PlastReconstr Surg , 1999 , 103 (1) : 1.
47. Hasan Q , Tan ST , Gush J , et al. Steroid therapy of a proliferating hemangioma : histochemical and molecular changes[J ] . Pediatrics , 2000 , 105 (1 Pt 1) : 117.
48.Pollman MJ , Naumovski L , Gibbons GH , et al. Vascular cellapoptosis :cell type - specific modulation by transforming growthfactor betal in endothelial cells versus smooth muscle cells[J ] .Circulation , 1999 , 99 (15) : 2019.
49.Hassn Q,Ruger BM,Mulliken JB,et al Clust\ ApoJ expression during the development of hemangioma Hum pathol,2003,31:691—697.
50.Tan ST,Wallis RA,He Y,et al Mast cells and hemangioma Plast Reconstr Surg,2004,113:999—1011.
51.曾嵘,李进. c - myc的结构和功能[J ] .国外医学:遗传学分册,1997 ,20(3) :116 - 119.
52. Willims GT ,Smith CA. Molecular regulation of apoptosis : geneticcontroloncell death[J ] . Cell ,1993 ,74 :777.
53.孙润芹,王康敏,黄莺,等。小儿皮肤血管瘤c-myc蛋白表达与其细胞增殖和凋亡的关系[J]。实用儿科临床杂志,2005,2(2):147—149.
54. Lee MH, Murphy G. Matrix metalloproteinases at a glance. J Cell Sci2004; 117(Pt 18): 4015-4016.
55. Abilleira S, Bevan S, Markus HS. The role of genetic variants of matrixmetalloproteinases in coronary and carotid atheroscleros is . J Med Genet2006; 43(12): 897-901.
56. Takahashi K, . Mulliken JB , Kozakewich HPW, et al. CellularMarkers That Distinguish the Phases of hemangioma during In2fancy and Childhood[J].JClin Invest ,1994 ,93 :2357-2364.
57. Bressler R ,Johnson DG. Pharmacological regulation of bloodglucose levels in noninsulin-dependent diabetes mellitus [J] .Arch Intern Med ,1997 ,157 (8) :836-848.
58.王玉红,牛爱国,徐晓青,等。血管内皮生长因子与基质金属蛋白酶2在血管瘤不同生长过程中的表达特征[J].中国组织工程研究与临床康复,2007,6(23):4488—4491.
59. Sun Y,Lin R, Dai J, et al. Supp ression of tumor growth using atisenseoligonucleotide against survivin in an orthotop ic transp lantmodel of human hepatocellular carcinoma in nude mice [ J ]. O ligonucleotides,2006, 16 (4) : 365 - 374.